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Label-Free and Simultaneous Mechanical and Electrical Characterization of Single Plant Cells Using Microfluidic Impedance Flow Cytometry

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Abstract

Despite that single-cell-type-level analyses have been extensively conducted on animal models to gain new insight into complex biological processes, the unique biological and physiological properties of plant cells have not been widely studied at single-cell resolution. In this work, an electrical impedance flow cytometry was fabricated based on microfluidics with constriction microchannel to simultaneously characterize the mechanical and electrical properties of single plant cells. Protoplasts from two model plant species, the herbaceous Arabidopsis thaliana and the woody Populus trichocarpa, could be readily discriminated by their respective mechanical traits, but not by electrical impedance. On the contrary, overexpression of a red fluorescent protein (RFP) on plasma membrane resulted in changes in cell electrical impedance instead of cell deformability. During primary cell wall (PCW) regeneration, this extracellular layer outside of protoplasts introduced dramatic variations in both mechanical and electrical properties of single plant cells. Furthermore, the effects of auxin, an essential phytohormone regulating PCW reformation, were validated on this platform. Taken together, our results revealed a novel application of microfluidic impedance flow cytometry in the field of plant science to simultaneously characterize dual biophysical properties at single cell resolution, which could be further developed as a powerful and reliable tool for plant cell phenotyping and cell fate specification.

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... Constriction channel allows for more accurate detection of specific membrane capacitance and cytoplasm conductivity, thus increasing the classification success rate of different cell populations [58]. Hence, it has been used for screening various types of single cells, such as blood cells [59], tumor cells [43,58] and plant cells [60]. Due to the cross-sectional area of the constriction channel must be smaller than the size of the interested cells, this design has higher risk of channel blockage and lower throughput. ...
... The passage time for cells to pass through the constriction channel is related to cellular mechanical properties. In order to obtain the passage time, Han et al. introduced a pairs of coplanar electrodes at the inlet and outlet of the constriction channel [60]. The time point when a cell passes through each sensing unit is recorded, and then the passage time can be calculated according to the time interval for a cell from a sensing unit to another. ...
... Recent IFC devices applied in single-cell analysis are summarized in Table 1. These applications, discussed in this subsection, are simply classified according to cell species, including blood cells [110][111][112][113][114], tumor cells [43,52,[115][116][117][118][119][120][121][122], stem cells [123][124][125][126][127], plant cells [60,[128][129][130][131][132] and microbes [53,62,[133][134][135][136][137][138][139][140][141]. In terms of blood cells, researchers focused on the identification and counting of normal or diseased blood cells. ...
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As stem cell therapies become more common in the clinic, there is a greater need for real-time, label-free monitoring of the differentiation status of the cells. In this paper, we present a dual-micropore-based, high-throughput microfluidic electrical impedance flow cytometer for non-invasive identification of the differentiation state of mesenchymal stem cells. The mesenchymal stem cells were induced to differentiate into osteoblasts over a 21 day period. Samples of mesenchymal stem cells and osteoblasts were flowed through the device, and impedance measurements were acquired over a frequency range from 50 kHz to 10 MHz. The opacity and relative angle, which shed light on the membrane capacitance and interior dielectric properties of cells, were used as interrogation parameters to analyze collected impedance data. Specifically, identification of mesenchymal stem cells and osteoblasts in a mixed population was optimized using a combination of opacity signature at 500 kHz and relative angle at 3 MHz. Identification of both cell populations in a mixed sample was successfully achieved with an accuracy of 87%. The results show a progressive increase in the number of osteoblasts throughout the 21 day differentiation process, with 36% more mesenchymal stem cells differentiated after 14 days of induction compared to after just 7 days. The dual-micropore microfluidic impedance flow cytometer system may become an important non-invasive tool for assessing stem cell quality and differentiation stages for future regenerative medicine applications.
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We developed a microfluidic sensor for label-free flow cytometric cell differentiation by combined multiple AC electrical impedance and light scattering analysis. The measured signals are correlated to cell volume, membrane capacity and optical properties of single cells. For an improved signal to noise ratio, the microfluidic sensor incorporates two electrode pairs for differential impedance detection. One-dimensional sheath flow focusing was implemented, which allows single particle analysis at kHz count rates. Various monodisperse particles and differentiation of leukocytes in haemolysed samples served to benchmark the microdevice applying combined AC impedance and side scatter analyses. In what follows, we demonstrate that AC impedance measurements at selected frequencies allow label-free discrimination of platelets, erythrocytes, monocytes, granulocytes and lymphocytes in whole blood samples involving dilution only. Immunofluorescence staining was applied to validate the results of the label-free cell analysis. Reliable differentiation and enumeration of cells in whole blood by AC impedance detection have the potential to support medical diagnosis for patients with haemolysis resistant erythrocytes or abnormally sensitive leucocytes, i.e. for patients suffering from anaemia or leukaemia.
Article
The advent of user-friendly instruments for measuring force/deflection curves of plant surfaces at high spatial resolution has resulted in a recent outpouring of reports of the 'Young's modulus' of plant cell walls. The stimulus for these mechanical measurements comes from biomechanical models of morphogenesis of meristems and other tissues, as well as single cells, in which cell wall stress feeds back to regulate microtubule organization, auxin transport, cellulose deposition, and future growth directionality. In this article I review the differences between elastic modulus and wall extensibility in the context of cell growth. Some of the inherent complexities, assumptions, and potential pitfalls in the interpretation of indentation force/deflection curves are discussed. Reported values of elastic moduli from surface indentation measurements appear to be 10- to >1000-fold smaller than realistic tensile elastic moduli in the plane of plant cell walls. Potential reasons for this disparity are discussed, but further work is needed to make sense of the huge range in reported values. The significance of wall stress relaxation for growth is reviewed and connected to recent advances and remaining enigmas in our concepts of how cellulose, hemicellulose, and pectins are assembled to make an extensible cell wall. A comparison of the loosening action of α-expansin and Cel12A endoglucanase is used to illustrate two different ways in which cell walls may be made more extensible and the divergent effects on wall mechanics.
Article
The dielectric properties of tumour cells are known to differ from normal blood cells, and this difference can be exploited for label-free separation of cells. Conventional measurement techniques are slow and cannot identify rare circulating tumour cells (CTCs) in a realistic timeframe. We use high throughput single cell microfluidic impedance cytometry to measure the dielectric properties of the MCF7 tumour cell line (representative of CTCs), both as pure populations and mixed with whole blood. The data show that the MCF7 cells have a large membrane capacitance and size, enabling clear discrimination from all other leukocytes. Impedance analysis is used to follow changes in cell viability when cells are kept in suspension, a process which can be understood from modelling time-dependent changes in the dielectric properties (predominantly membrane conductivity) of the cells. Impedance cytometry is used to enumerate low numbers of MCF7 cells spiked into whole blood. Chemical lysis is commonly used to remove the abundant erythrocytes, and it is shown that this process does not alter the MCF7 cell count or change their dielectric properties. Combining impedance cytometry with magnetic bead based antibody enrichment enables MCF7 cells to be detected down to 100 MCF7 cells in 1 ml whole blood, a log 3.5 enrichment and a mean recovery of 92%. Microfluidic impedance cytometry could be easily integrated within complex cell separation systems for identification and enumeration of specific cell types, providing a fast in-line single cell characterisation method.
Article
We present a novel Multi-Regime Analysis (MRA) routine for interpreting force indentation measurements of soft materials using atomic force microscopy. The MRA approach combines both well established and semi-empirical theories of contact mechanics within a single framework to deconvolute highly complex and non-linear force-indentation curves. The fundamental assumption in the present form of the model is that each structural contribution to the mechanical response acts in series with other ‘mechanical resistors’. This simplification enables interpretation of the micromechanical properties of materials with hierarchical structures and it allows automated processing of large data sets, which is particularly indispensable for biological systems. We validate the algorithm by demonstrating for the first time that the elastic modulus of polydimethylsiloxane (PDMS) films is accurately predicted from both approach and retraction branches of force-indentation curves. For biological systems with complex hierarchical structures, we show the unique capability of MRA to map the micromechanics of live plant cells, revealing an intricate sequence of mechanical deformations resolved with precision that is unattainable using conventional methods of analysis. We recommend the routine use of MRA to interpret AFM force-indentation measurements for other complex soft materials including mammalian cells, bacteria and nanomaterials.
Article
Plant and animals have evolved different strategies for their development. Whether this is linked to major differ-ences in their cell mechanics remains unclear, mainly because measurements on plant and animal cells relied on independent experiments and setups, thus hindering any direct comparison. In this study we used the same micro-rheometer to compare animal and plant single cell rheology. We found that wall-less plant cells exhibit the same weak power law rheology as animal cells, with comparable values of elastic and loss moduli. Remarkably, microtubules primarily contributed to the rheological behavior of wall-less plant cells whereas rheology of animal cells was mainly dependent on the actin network. Thus, plant and animal cells evolved different molecular strategies to reach a comparable cytoplasmic mechanical core, suggesting that evolutionary convergence could include the internal biophysical properties of cells.
Article
Conventional methods of plant cell analysis rely on growing plant cells in soil pots or agarose plates, followed by screening the plant phenotypes in traditional greenhouses and growth chambers. These methods are usually costly, need a large number of experiments, suffer from low spatial resolution and disorderly growth behavior of plant cells, with lack of ability to locally and accurately manipulate the plant cells. Microfluidic platforms take advantage of miniaturization for handling small volume of liquids and providing a closed environment, with the purpose of in vitro single cell analysis and characterizing cell response to external cues. These platforms have shown their ability for high-throughput cellular analysis with increased accuracy of experiments, reduced cost and experimental times, versatility in design, ability for large-scale and combinatorial screening, and integration with other miniaturized sensors. Despite extensive research on animal cells within microfluidic environments for high-throughput sorting, manipulation and phenotyping studies, the application of microfluidics for plant cells studies has not been accomplished yet. Novel devices such as RootChip, RootArray, TipChip, and PlantChip developed for plant cells analysis, with high spatial resolution on a micrometer scale mimicking the internal microenvironment of plant cells, offering preliminary results on the capability of microfluidics to conquer the constraints of conventional methods. These devices have been used to study different aspects of plant cell biology such as gene expression, cell biomechanics, cellular mechanism of growth, cell division, and cells fusion. This review emphasizes the advantages of current microfluidic systems for plant science studies, and discusses future prospects of microfluidic platforms for characterizing plant cells response to diverse external cues.
Article
A method is described for purifying plant protoplasts from cellular and subcellular debris. The procedure utilizes a density buffer containing 9.6% sodium metrizoate and 5.6% Ficoll. The use of fluorescein diacetate for assessing the viability of plant protoplasts is also reported.
Article
Tip-growing cells have the unique property of invading living tissues and abiotic growth matrices. To do so, they exert significant penetrative forces. In plant and fungal cells, these forces are generated by the hydrostatic turgor pressure. Using the TipChip, a microfluidic lab-on-a-chip device developed for tip-growing cells, we tested the ability to exert penetrative forces generated in pollen tubes, the fastest-growing plant cells. The tubes were guided to grow through microscopic gaps made of elastic polydimethylsiloxane material. Based on the deformation of the gaps, the force exerted by the elongating tubes to permit passage was determined using finite element methods. The data revealed that increasing mechanical impedance was met by the pollen tubes through modulation of the cell wall compliance and, thus, a change in the force acting on the obstacle. Tubes that successfully passed a narrow gap frequently burst, raising questions about the sperm discharge mechanism in the flowering plants.
Article
Biomechanical and mathematical modeling of plant developmental processes requires quantitative information about the structural and mechanical properties of living cells, tissues and cellular components. A crucial mechanical property of plant cells is the mechanical stiffness or Young's modulus of its cell wall. Measuring this property in situ at single cell wall level is technically challenging. Here, a bending test is implemented in a chip, called Bending-Lab-On-a-Chip (BLOC), to quantify this biomechanical property for a widely investigated cellular model system, the pollen tube. Pollen along with culture medium is introduced into a microfluidic chip and the growing pollen tube is exposed to a bending force created through fluid loading. The flexural rigidity of the pollen tube and the Young's modulus of the cell wall are estimated through finite element modeling of the observed fluid-structure interaction. An average value of 350 MPa was experimentally estimated for the Young's modulus in longitudinal direction of the cell wall of Camellia pollen tubes. This value is in agreement with the result of an independent method based on cellular shrinkage after plasmolysis and with the mechanical properties of in vitro reconstituted cellulose-callose material.
Article
Large scale phenotyping of tip growing cells such as pollen tubes has hitherto been limited to very crude parameters such as germination percentage and velocity of growth. To enable efficient and high throughput execution of more sophisticated assays, an experimental platform was developed based on microfluidic and MEMS (microelectromechanical systems) technology, the TipChip. The device allows positioning of pollen grains or fungal spores at the entrances of serially arranged microchannels harboring microscopic experimental setups. The tip growing cells, pollen tubes, filamentous yeast or fungal hyphae, can be exposed to chemical gradients, microstructural features, integrated biosensors or directional triggers within the modular microchannels. The device is compatible with Nomarski optics and fluorescence microscopy. Using this platform we were able to answer several outstanding questions on pollen tube growth. We established that unlike root hairs and fungal hyphae, pollen tubes do not have a directional memory. Furthermore, pollen tubes were found to be able to elongate in air raising the question how and where water is taken up by the cell. The platform opens new avenues for both, more efficient experimentation and large scale phenotyping of tip growing cells under precisely controlled, reproducible conditions. © 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd.
Article
The phytohormone auxin is a primary regulator of growth and developmental pattern formation in plants. Auxin accumulates at specific sites (e.g., organ primordia) and induces localized growth within a tissue. Auxin also mediates developmental responses to intrinsic and external physical stimuli; however, exactly how mechanics influences auxin distribution is unknown. Here we show that mechanical strain can regulate auxin transport and accumulation in the tomato shoot apex, where new leaves emerge and rapidly grow. Modification of turgor pressure, application of external force, and artificial growth induction collectively show that the amount and intracellular localization of the auxin efflux carrier PIN1 are sensitive to mechanical alterations. In general, the more strained the tissue was, the more PIN1 was present per cell and the higher the proportion localized to the plasma membrane. Modulation of the membrane properties alone was sufficient to explain most of the mechanical effects. Our experiments support the hypothesis that the plasma membrane acts as a sensor of tissue mechanics that translates the cell wall strain into cellular responses, such as the intracellular localization of membrane-embedded proteins. One implication of this fundamental mechanism is the mechanical enhancement of auxin-mediated growth in young organ primordia. We propose that growth-induced mechanical strain upregulates PIN1 function and auxin accumulation, thereby promoting further growth, in a robust positive feedback loop.
Article
Mechanical properties of cells have been shown to have a significant role in disease, as in many instances cell stiffness changes when a cell is no longer healthy. We present a high-throughput microfluidics-based approach that exploits the connection between travel time of a cell through a narrow passage and cell stiffness. The system resolves both cell travel time and relative cell diameter while retaining information on the cell level. We show that stiffer cells have longer transit times than less stiff ones and that cell size significantly influences travel times. Experiments with untreated HeLa cells and cells made compliant with latrunculin A and cytochalasin B further demonstrate that travel time is influenced by cell stiffness, with the compliant cells having faster transit time.
Article
1. Sections of auxin-starved hypocotyls of Helianthus annuus do not show any significant growth rate in water of buffers of pH\>-6. However, in buffers with pH-values of approximately 4, elongation growth is observed; its rate is similar to the rate of auxin-stimulated growth (after 6 h incubation). \3- This phenomenon of acid-induced growth occurs also under anaerobic conditions in contrast to auxin-induced growth (Hager 1962). 2. Intact cell wall aggregates of Helianthus hypocotyls were obtained by complete plasmolysis of hypocotyls in 50% glycerol; cell wall associated enzymes were still active after this treatment, at least in part. While cell walls in solutions of pH\>-6 show only a small plastic extension during the first minute in response to a 50 g stretching force, a constant rate of elongation over longer periods of time (measured up to 1 h) is observed in weakly acid buffers. The highest rate of elongation is observed at about pH 4. This acid-induced plastic extension is completely inhibited by Cu2+-ions (5mM); the elongation of cell walls is apparently the consequence of an enzyme-catalysed increase in plasticity having a pH optimum of about 4. The pH optimum of acid-induced cell wall extension observed during stretching of plasmolysed hypocotyls coincides with the pH optimum of acid-induced growth of intact hypocotyl sections (around pH 4). 3. Under anaerobic conditions the growth rate of intact coleoptiles stays unchanged (at a low value) if the sections are incubated in a buffer of pH 5.0. Higher proton concentrations, however, stimulate growth immediately, whereas low proton concentrations are inhibitory (Fig. 7 and 8). The strongest initial growth response is elicited by buffers or acids of pH 3.9 (Fig. 9). Acid-induced growth of coleoptiles with a similar pH optimum is also found under anaerobic conditions. The growth of coleoptile cylinders can be switched on or off by repeatedly changing to acid or basic medium, respectively (under conditions of anaerobiosis) (Fig. 10). IAA-induced growth (aerobic conditions, pH5) can also be inhibited immediately by basic buffers or NaOH-solutions, and resumes after the pH value is lowered (Fig. 11). This pH-dependency may be taken as an indication that auxin affects the same reaction which is stimulated by high proton concentrations and which may be the last step in the process of cell elongation. CCCP, known to make membranes permeable for protons, rapidly inhibits the auxin-induced elongation growth (pH 6,5) when applied at a concentration which does not influence respiration; removal of CCCP shows that the growth inhibition by CCCp is partly reversible (Fig. 12). In contrast, acid-induced elongation growth (pH 4) shows inhibition by CCCP not before 10 min after application.—These findings suggest that auxin induces a proton accumulation in a cell wall compartment and, as a consequence, enzymatic cell wall softening. Such an accumulation could be brought about by an auxin-activated, membrane-bound, anisotropic ATPase or ion pump. The notion that ATPases or pumps may be located in the outer layers of the cell membrane is supported by the observation that addition of ATP to coleoptile cylinders under anaerobic conditions results in an immediate stimulation of elongation (Fig. 14). This effect is further enhanced by Mg2+-and K+-ions (Figs. 15 and 16). Mg2+ can be partly replaced by Ca2+. The stimulatory effect of ATP is increased considerably if the coleoptiles are treated with IAA under aerobic conditions prior to ATP addition (Figs. 15 b and 14). ITP, GTP, UTP, and CTP induce elongation growth under anaerobiosis similarly to ATP. In the presence of ITP or GTP the increase in growth rate is maintained over a longer period of time than in the presence of the other nucleoside triphosphates (Fig. 17). IAA, which causes no elongation growth under anaerobiosis (Fig. 13) is also unable to further stimulate the elongation growth induced by ATP, UTP, or CTP under anaerobiosis (Fig. 18); however, if IAA is added after growth has been stimulated by GTP or ITP, a temporary inhibition and, 10 min later, a strong stimulation is noticed (Fig. 19). If the sequence of addition is reversed, —that is, if IAA (without growth effect) and, after 20 min, GTP or ITP are added to the coleoptiles—, the same initial inhibition and subsequent increase of the growth rate is found (Fig. 20). Thus, IAA can stimulate growth of coleoptiles even under anaerobic conditions if GTP or ITP is present at the same time. 4. The results support the following hypothesis (Fig. 21): auxin acts cooperatively with GTP (ITP) as an effector of a membrane-bound, anisotropic ATPase or proton pump. This pump, activated by auxin, utilizes respiration energy (ATP or other nucleoside triphosphates) to raise the proton concentration in a compartment at the cell wall. This event leads to an increase in the activity of enzymes softening cell walls and thus triggers cell elongation. The transport or secretion of protons into the cell wall compartment should be compensated by a flow of cations into the interior of the cytoplasm or by a flow of anions to the cell periphery, thus causing secondary auxin effects.
Article
The shape of an organism relies on a complex network of genetic regulations and on the homeostasis and distribution of growth factors. In parallel to the molecular control of growth, shape changes also involve major changes in structure, which by definition depend on the laws of mechanics. Thus, to understand morphogenesis, scientists have turned to interdisciplinary approaches associating biology and physics to investigate the contribution of mechanical forces in morphogenesis, sometimes re-examining theoretical concepts that were laid out by early physiologists. Major advances in the field have notably been possible thanks to the development of computer simulations and live quantitative imaging protocols in recent years. Here, we present the mechanical basis of shape changes in plants, focusing our discussion on undifferentiated tissues. How can growth be translated into a quantified geometrical output? What is the mechanical basis of cell and tissue growth? What is the contribution of mechanical forces in patterning?
Article
Microfabricated flow cytometers can detect, count, and analyze cells or particles using microfluidics and electronics to give impedance-based characterization. Such systems are being developed to provide simple, low-cost, label-free, and portable solutions for cell analysis. Recent work using microfabricated systems has demonstrated the capability to analyze micro-organisms, erythrocytes, leukocytes, and animal and human cell lines. Multifrequency impedance measurements can give multiparametric, high-content data that can be used to distinguish cell types. New combinations of microfluidic sample handling design and microscale flow phenomena have been used to focus and position cells within the channel for improved sensitivity. Robust designs will enable focusing at high flowrates while reducing requirements for control over multiple sample and sheath flows. Although microfluidic impedance-based flow cytometers have not yet or may never reach the extremely high throughput of conventional flow cytometers, the advantages of portability, simplicity, and ability to analyze single cells in small populations are, nevertheless, where chip-based cytometry can make a large impact.
Article
Plants are under tremendous mechanical forces generated by turgor pressure. How do these forces mediate growth and development? In order to answer this question, it is necessary to understand the mechanics of growth and morphogenesis. In this 'mathless' tutorial, the concepts of strain, mechanical stress and buckling are reviewed and illustrated with recent work on leaf shape, on leaf vasculature, and on organogenesis at the shoot apical meristem.
Article
Many genes and molecules that drive tissue patterning during organogenesis and tissue regeneration have been discovered. Yet, we still lack a full understanding of how these chemical cues induce the formation of living tissues with their unique shapes and material properties. Here, we review work based on the convergence of physics, engineering and biology that suggests that mechanical forces generated by living cells are as crucial as genes and chemical signals for the control of embryological development, morphogenesis and tissue patterning.
Article
Miniature high speed label-free cell analysis systems have yet to be developed, but have the potential to deliver fast, inexpensive and simple full blood cell analysis systems that could be used routinely in clinical practice. We demonstrate a microfluidic single cell impedance cytometer that performs a white blood cell differential count. The device consists of a microfluidic chip with micro-electrodes that measure the impedance of single cells at two frequencies. Human blood, treated with saponin/formic acid to lyse erythrocytes, flows through the device and a complete blood count is performed in a few minutes. Verification of cell dielectric parameters was performed by simultaneously measuring fluorescence from CD antibody-conjugated cells. This enabled direct correlation of impedance signals from individual cells with phenotype. Tests with patient samples showed 95% correlation against commercial (optical/Coulter) blood analysis equipment, demonstrating the potential clinical utility of the impedance microcytometer for a point-of-care blood analysis system.
Article
Mechanotransduction research has focused historically on how externally applied forces can affect cell signalling and function. A growing body of evidence suggests that contractile forces that are generated internally by the actomyosin cytoskeleton are also important in regulating cell behaviour, and suggest a broader role for mechanotransduction in biology. Although the molecular basis for these cellular forces in mechanotransduction is being pursued in cell culture, researchers are also beginning to appreciate their contribution to in vivo developmental processes. Here, we examine the role for mechanical forces and contractility in regulating cell and tissue structure and function during development.
Article
Plant cells encase themselves within a complex polysaccharide wall, which constitutes the raw material that is used to manufacture textiles, paper, lumber, films, thickeners and other products. The plant cell wall is also the primary source of cellulose, the most abundant and useful biopolymer on the Earth. The cell wall not only strengthens the plant body, but also has key roles in plant growth, cell differentiation, intercellular communication, water movement and defence. Recent discoveries have uncovered how plant cells synthesize wall polysaccharides, assemble them into a strong fibrous network and regulate wall expansion during cell growth.
Article
Direct measurements of the volumetric elastic modulus, in, of cells of a higher plant were performed on the epidermal bladder cells of Mesembryanthemum crystallinum using a pressure probe technique. Measurements on giant algal cells (Valonia, Nitellopsis) are given for comparison. Giant celled algae and M. crystallinum bladders have elastic moduli, in, which depend strongly on turgor pressure, P, and on cell volume, V. The in values of Mesembryanthemum bladders range between 5 bar at zero pressure and 100 bar at full turgor pressure (3-4 bar). in increased with cell size (volume) at a given turgor pressure, and this volume dependence was pronounced more in the high pressure range. From the in (P) characteristics, complete volume-pressure curves were obtained for Mesembryanthemum bladders and giant algal cells. The results suggest that the in (P) and in (V) characteristics of all plant cells are similar. The significance of the pressure and volume effects for the water relations and growth processes of plant cells is discussed briefly.
Article
The manipulation of fluids in channels with dimensions of tens of micrometres--microfluidics--has emerged as a distinct new field. Microfluidics has the potential to influence subject areas from chemical synthesis and biological analysis to optics and information technology. But the field is still at an early stage of development. Even as the basic science and technological demonstrations develop, other problems must be addressed: choosing and focusing on initial applications, and developing strategies to complete the cycle of development, including commercialization. The solutions to these problems will require imagination and ingenuity.